School of Bioprocess Engineering

ERT 317/4
Biochemical Engineering
Sem 1 2015/2016

TUTORIAL 1 (Ans Scheme)

1. An enzymatic assay was carried under two different sets of conditions out using a pure
substrate S. The results are tabulated below (Table 1).
Table 1
[S]/ Vo
10-5 M Condition A Condition B
1.5 0.21 0.08
2.0 0.25 0.10
3.0 0.28 0.12
4.0 0.33 0.13
8.0 0.44 0.16
16.0 0.40 0.18

a) Plot the data using the Lineweaver-Burke plot

For Lineweaver-Burke Plot

 b)

c) Calculate the values of Vmax and Km for both sets of conditions

Cond A Cond B

2. Demonstrate a possible scheme that may be used to describe pH dependence of the

enzymatic reaction rate for ionizing enzymes. Explain how ionic groups at the active site
of an enzyme may result in a variation of activity with pH.
 -Enzyme are active only over small range of pH due to the active site functional group
charges (ionic form) and the three dimensional shape of enzyme are pH-dependent
-Certain enzyme have ionic group on their active sites, and these ionic group must be in a
suitable form (acid or base) to function.
-Variation in pH of medium result in changes of:
-Ionic form of the active site
-Activity of enzyme, hence the reaction rate
-Affect the maximum reaction rate, Km
-stability of enzyme

3. Define the immobilized enzyme systems.

Restriction of enzyme mobility in a fixed space

4. Classify the methods of immobilized soluble enzymes by the aid of a diagram.

5. What type of inhibitor should be useful in trying to identify what features of a substrate
are important for binding the substrate to the active site of an enzyme catalyzing a single-
substrate reaction? Name that related method of immobilization?

Competitive inhibitor
Surface immobilization: Covalent bonding